Tuberculosis is a chronic infectious diseases caused by Mycobacterium tuberculosis(MTB).As the drug-resistance characteristics are different in patients with various genotypes,thus,the gene polymorphism study have critical clinical significance.Among the all kinds of techniques,some have been used to analyze polymorphism for a long time and new development in that respect has also been made recently.On the other hand,some techniques are e-merging but demonstrate promising application prospects.This study summarizes the gene polymorphism study of MTB which have been used or are emerging in recent years,and points out a few shortcomings briefly.Our object is to make a contribution to theoretical basis and knowledge accumulation in the drug-resistance and epidemiological survey field.

Objective To investigate the effect of CXCL16 on the development of murine collageninduced arthritis (CIA). Methods CXCL16 mRNA of the involved synovium and serum CXCL16 protein were determined respectively by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA) in murine collagen-induced arthritis. The proliferation of lymphocytes from murine spleen and the level of RANKL mRNA, stimulated by CXCL16 at different concentrations (0,100, 200, 400, 800 ng/ml), was detected respectively by CCK-8 and RT-PCR, then the level of IL-2 and IFN-γ in culture supernatant was detected by ELISA. Comparisons between groups were tested by t test and one-way ANOVA analysis. Results The serum CXCL16 [(127± 10) vs (72±8) pg/ml, P＜0.05] and synovial CXCL16 mRNA (0.214±0.007 vs 0.375±0.009, P＜0.01) in CIA were all significantly higher than those in normal controls. The proliferation of CXCL16 (200, 400, 800 ng/ml) in CIA mouse lymphocytes, was significantly higher than that of CXCL16 (0 ng/ml) (0.51±0.06, 0.56±0.05, 0.55±0.04, (0.41±0.04, P＜0.05). And CXCL16 on the CIA stimulated lymphocyte proliferation was significantly higher than controls on normal lymphocytes (P＜0.05). Compared with blank control group, the expression of IL-2, IFN-γ and RANKL mRNA of CIA CXCL16 (400, 800 ng/ml) groups was higher significantly (P＜0.01). Conclusion CXCL16 plays an important role in the development of murine CIA by activating lymphocytes.

Objective To detect D-Dimer in the blood of rheumatoid arthritis (RA) patients and to investigate its clinical significance in RA. Methods Blood samples were obtained from 58 patients with RA,18 patients with systemic lupus erythematosus (SLE), 15 patients with ankylosing spondylitis, 11 patients with osteoarthritis, and 20 patients with other connective tissue diseases. The presence of thrombotic diseases was excluded in all patients. The presence of D-Dimer in the blood was examined by immunoturbidimetry. The following clinical and laboratory data were collected: disease activity index DAS28, rheumatoid factor (RF),erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP). Mann-Whitney U test, t-test ,Spearman correlation were used for statistical analysis. Results ① The prevalence of high level D-Dimer in RA patients was higher than that of the control group (82.6％ vs 21.9％, P＜0.01). The titer of D-Dimer in RA was significantly higher than that of the control group [(1.76±1.57) vs (0.32±0.25) mg/L, P＜0.01]. ②DAS28 was higher in RA patients with positive D-Dimer than those with negative D-Dimer (5.4±1.0 vs 4.4±0.8,P＜0.01). The values of ESR, CRP and RF in RA patients with positive D-Dimer were significantly higher than those in patients with negative D-Dimer (P＜0.05). ③ There was positive correlation between D-Dimer and DAS28 (r=0.406, P＜0.05), ESR (r=0.355, P＜0.01), and RF (r=0.319, P＜0.05). Conclusion The level of D-Dimer in the blood of RA patients is significantly higher than other rheumatic diseases, and is positively correlated with disease activities. The results indicated that the activation of coagulation-fibrinolytic systems may play a role in the pathogenesis of rheumatoid arthritis.

AIM:To determine which species of snake venoms contained protein c activator among 9 species of Chinese snake venoms. METHODS: Anticoagulant activity was examined by activated partial thromboplastin time (APTT) assay，and amidolytic activity was measured with activated protein c (APC) specific chromogenic peptide substrate-chromozy APC. RESULTS: Among 9 species of Chinese snake venoms，Trimeresurus mucrosquamatus venom and Agkistrodon halys venom were not only able to generate amidolytic activity from purified human PC, but also prolonged APTT strongly even at such a concentration as 1.5 mg/L.CONCLUSION: Trimeresurus mucrosquamatus venom and Agkistrodon halys venom contain protein c activator which activating human plasma PC into APC.

Objective To investigate the clinical features and risk factors of spondyloarthropathy with anterior uveitis. Methods The clinical and laboratory data of 86 patients with spondyoarthropathy associated with anterior uveitis in our hospital were collected, analyzed and summarized from March 2005 to December 2008, and the patients were followed up as closely as possible. The data of the 86 patients were compared with 93 patients who had spondyloarthropathy without anterior uveitis at the same period. All data were analyzed by using SPSS11.5 software package. Results Compared with non-ophthalmia group, ophthalmia group had significantly longer course[(11 ±8)vs(5±6), P＜0.01], and higher proportion of positive family history(27.9%vs 9.7%, P＜0.01), the proportion of low back pain at night, morning stiffness, spinal deformity, limitation of waist-bending and severe sacroiliac joint lesions were all significantly higher(P＜0.05), HLA-B27 positive rate was significantly higher as well(92.2% vs 81.5%, P＜0.05). The attack of uveitis usually had seasonality and precipitating cause. The patients with anterior uveitis as first symptom had significant higher frequency of ophthalmitis(P＜0.01), the ratio of eye permanent lesions was also significantly higher(P＜0.01). The frequencies of attack were positively correlated with the course of disease(r=0.294, P=0.006), Logistic multiple regression analysis showed that the incidence risk of ophthalmia were related to the course of disease(P=0.013, OR=1.099, 95%CI 1.030～1.183)and severe sacroiliac joint lesions(P=0.012, OR=3.071, 95%CI 1.286 ～7.314). Conclusion The spondyloarthropathy associated with anterior uveitis had its own characteristics, We should pay attention to the risk factors of anterior uveitis,and prevent the recurrence of ophthalmia.

Essential tremor (ET) is a common movement disorder. It is characterized by a distinctive 4-12 Hz action tremor typically affecting bilateral upper limbs. Existing drugs for ET mainly include β-blockers, anticonvulsants, benzodiazepines, etc. However, the efficacy of existing drugs is limited. With the development of the medical research, some progress has been made in the treatment of ET. The review will explore the recent advances in the treatment of ET,such as new drugs, surgical treatment, repetitive transcranial magnetic stimulation, rehabilitation treatment, etc., in order to provide clinical application prospects.

Objective To detect influenza A virus by reverse transcription‐loop mediated isothermal amplification (RT‐LAMP) assay to established a rapid ,simple and visualization nucleic acid detection method .Methods The RT‐LAMP primers were designed in accordance with the hemagglutinin gene of influenza A virus .Then ,the specificity of the primers was evaluated by detection of different influenza viruses ,and the sensitivity was confirmed by testing multiple diluted RNA samples . Hydroxynaphthol blue (HNB) was used for visually evaluation and gel electrophoresis was used for validation .Clinical samples were detected by RT‐LAMP assay .Its consistency with fluorescent quantitative polymerase chain reaction (PCR) methods was compared .Results The primers of RT‐LAMP assay had high specificity . This technique could amplify influenza A virus accurately .The detection limit of RT‐LAMP assay was 2 .5 × 103 copies/mL by detection of multiple diluted RNA samples .In addition ,the results of RT‐LAM P assay could be visually inspected using HNB by color change ,and the results was in accordance with that of gel electrophoresis . RT‐LAMP assay was in consistence with fluorescent quantitative PCR when clinically applied .Conclusions RT‐LAMP assay is a rapid ,specific ,sensitive and simple method to detect influenza A virus .

Objective:To clone and express the Der f 7 recombinant protein from Dermatophagoides farinae and prepare the Der f 7 monoclonal antibody.Methods: The Der f 7 recombinant protein was expressed in prokaryotic expression system of pET28a(+)-Der f 7.BALB /c mice were immunized with the recombinant protein.Myeloma cells and spleen cells were fused,and hybridoma cells were screened by ELISA.Hybridoma cells were injected into the mice abdominal cavity to obtain ascites.It was purified by protein A agarose medium ascites,and then to dentified the titer and purity of the antibody.The specificity of the antibody was identified by Western blot.Results: Three hybridoma cells which stably secret recombinant Der f 7 monoclonal antibody were obtained.The monoclonal antibody had high purity,the titer was higher than 1∶243 000.Western blot showed that Der f 7 recombinant protein could be recognized well.Conclusion: We successfully obtained Der f 7 monoclonal antibody,which can be used for the quantification and localization of Der f 7 allergen and the diagnosis and treatment of allergic diseases.

AIM:To determine which species of snake venoms contained protein c activator among 9 species of Chinese snake venoms. METHODS: Anticoagulant activity was examined by activated partial thromboplastin time (APTT) assay,and amidolytic activity was measured with activated protein c (APC) specific chromogenic peptide substrate-chromozy APC. RESULTS: Among 9 species of Chinese snake venoms,Trimeresurus mucrosquamatus venom and Agkistrodon halys venom were not only able to generate amidolytic activity from purified human PC, but also prolonged APTT strongly even at such a concentration as 1.5 mg/L.CONCLUSION: Trimeresurus mucrosquamatus venom and Agkistrodon halys venom contain protein c activator which activating human plasma PC into APC.

Objective To evaluate the prevalence and clinical significance of IgG,IgA and IgM isotypes of anti-peptidylarginine deiminase 4 (anti-PAD4) antibodies in early rheumatoid arthritis (RA).Methods IgG,IgA and IgM isotypes of anti-PAD4 antibodies were measured in the sera of 88 RA patients with disease duration less than 2 years,62 patients with other rheumatic diseases and 57 healthy subjects.The diagnostic performance of IgG,IgA and IgM isotypes of anti-PAD4 antibodies and their relationship with disease duration,DAS28,ESR,CRP,anti-CCP antibodies and rheumatoid factor (RF) were evaluated.Data analysis were performed using t test,U test and Spearman's association analysis.Results ① The sensitivities of IgG,IgA,and IgM isotypes for early RA were 28.41％,36.36％ and 9.09％ respectively.The specificities of IgG,IgA and IgM isotypes were 94.12％,93.28％ and 95.80％ respectively.② IgA isotype was positively correlated with age (r=0.234,P=0.028),DAS28 (r=0.309,P=0.007),ESR (r=0.382,P=0.000) and CRP (r=0.291,P=0.008),while negatively correlated with disease duration (r=-0.295,P=0.006).③ IgA isotype was positively correlated with IgG isotype (r=0.451,P＜0.01).In the IgG negative patients,the positivity of IgA isotype was 29％(18/63),which indicated that the IgA isotype might be helpful in diagnosing RA in IgG isotype negative patients.Conclusion Anti-PAD4 antibodies can be detected in early RA,primarily with IgA and IgG isotypes.IgA isotype has negative correlation with disease duration,indicating that IgA isotype of anti-PAD4 antibody may play a role in the very early stage RA.